The objective of our research proposal is to study the interaction of nonhistone chromosomal proteins (NHCP) with DNA/histone complexes and to chromatin. This involves the isolation of various NHCP fractions and the close examination of their binding to complexes of DNA/histones from urea-buffers. The complexes are later transcribed with homologous RNA polymerase B(II), and the RNA transcript analyzed for base sequence homology to in vivo poly A-messenger RNA, using the methods of kinetic-complexity analysis to generate a Crot curve by hybridization of RNA to cDNA. The cDNA is synthesized from poly A-mRNA isolated from mouse Ehrlich Ascites cells using RNA-dependent DNA polymerase from Avian myeloblastosis virus. Through such methods we can carry out analysis on the base sequences of the RNA being transcribed. In addition, we also examine which fraction of NHCP can be linked to synthesis of tissue specific RNA's. Our methods entail: (1) the isolation of chromatin from mouse Ehrlich ascites cells; (2) isolation of RNA polymerase from Ehrlich ascites cells; (3) fractionation of NHCP; (4) preparation of various DNA/chromosomal protein complexes; and (5) transcription. These types of experimentations will aid in determining the participation of nonhistones in the process of craniofacial development both in normal and malformed mouse embryos.